ABSTRACT Fluorescence guided surgery (FGS) is a growing field with increasing adoption, having about a dozen companies with FDA cleared systems for perfusion imaging with indocyanine green, and leading surgical device companies now acquiring such systems for marketing. Advanced molecular probes are being tested in hundreds of ongoing clinical trials in FGS, with a combination of research grade and modified commercial systems. However, each FGS system suffers from low dynamic range and high background signals from a range of sources. The C- MOLECULES camera has been systematically designed to solve these two key problems, by having high adaptive gain, inherently high digital bit conversion, and tight control over wavelength and temporal sampling. Suppression of tissue autofluorescence and room light background signals are maximized through narrow band filtering with each of two cameras, further processing the images for subtractive display. A unique patented technology for ultrafast time-gated intensified camera work is utilized to further maximize the fluorescence to ambient background, using specific detection that reaches the level of single photons per pixel. This system will be finalized in hardware and software with time-gated LED lamps for fluorophore activation. System comparison testing for the level of background suppression will be completed relative to other commercially available systems, with a goal of 10x increase in contrast through enhanced background suppression. Molecular image testing will utilize a fluorescent protein affibody that is being studied at microdoses, to image EGFR expression in human head and neck tumors. A confirmation of the ability to image fluorescent affibody at picomolar concentrations will be verified for maximal sensitivity. System development and testing is a collaborative project between DoseOptics and Dartmouth, each of which have outstanding expertise in development of the technology and pilot human trials, respectively. The team has a proven history of publishing their work, leading the field of FGS, and delivering products that are commercially translated.